Figure 7

Calcium dysregulation is central to grade 3 development.

(A) Heatmap of the calcium signalling pathway (dre04020) in control and the different grades of OPP, showing clear down-regulation of this pathway in grade 3 larvae. (B) Memantine, an antagonist of NMDA receptors, reduced the prevalence of the grade 3 phenotype dramatically [9 groups with 10–18 larvae (total larvae: 132) were analysed for 3 μΜ CPO and 9 groups with 13–25 larvae (total larvae: 200) for 3 μΜ CPO + 100 μΜ memantine; p < 0.001, Mann-Whitney U Statistic (T(9) = 126)]. Seven days post fertilization (dpf) larvae were pre-incubated with 100 μM memantine for 1 h followed by co-exposure with 100 μM memantine/3 μM CPO for 24 h. (C) BAPTA-AM, a permeable calcium chelator, also reduced the prevalence of the grade 3 phenotype significantly [7 groups with 12–42 larvae each (total larvae: 214) were analysed for 3 μΜ CPO and 5 groups with 11–18 larvae (total larvae: 70) for 3 μΜ CPO + 100 μΜ BAPTA-AM; p < 0.005, Student’s t-test (t(12) = 3.287)]. Six days post fertilization zebrafish larvae were pre-incubated with 100 μM BAPTA-AM for 24 h followed by 24 h co-exposure with 3 μM CPO.